Berberine Depresses Inflammation and Adjusts Smooth Muscle to Ameliorate Ulcerative Colitis of Cats by Regulating Gut Microbiota

ABSTRACT Intestinal microbiota dysbiosis is a well established characteristic of ulcerative colitis (UC). Regulating the gut microbiota is an effective UC treatment strategy. Berberine (BBR), an alkaloid extracted from several Chinese herbs, is a common traditional Chinese medicine. To establish the efficacy and mechanism of action of BBR, we constructed a UC model using healthy adult shorthair cats to conduct a systematic study of colonic tissue pathology, inflammatory factor expression, and gut microbiota structure. We investigated the therapeutic capacity of BBR for regulating the gut microbiota and thus work against UC in cats using 16S rRNA genes amplicon sequencing technology. Our results revealed that dextran sulfate sodium (DSS)-induced cat models of UC showed weight loss, diarrhea accompanied by mucous and blood, histological abnormalities, and shortening of the colon, all of which were significantly alleviated by supplementation with BBR. A 16S rRNA gene-based microbiota analysis demonstrated that BBR could significantly benefit gut microbiota. Western blot, quantitative PCR, and enzyme-linked immunosorbent assays (ELISAs) showed that in DSS-induced cat models, the expression of the inflammatory factors was increased, activating the JAK2/STAT3 signaling pathway, and treatment with BBR reversed this effect. The myosin light chain (MLC) phosphorylation in the smooth muscle of the intestines is associated with motility of inflammation-related diarrhea in cats. This study used gut flora analyses to demonstrate the anti-UC effects of BBR and its potential therapeutic mechanisms and offers novel insights into the prevention of inflammatory diseases using natural products. IMPORTANCE Ulcerative colitis (UC) is common in clinics. Intestinal microbiota disorder is correlated with ulcerative colitis. Although there are many studies on ulcerative colitis in rats, there are few studies on colitis in cats. Therefore, this study explored the possibility of the use of BBR as a safe and efficient treatment for colitis in cats. The results demonstrated the therapeutic effects of BBR on UC based on the state of the intestinal flora. The study found BBR supplementation to be effective against dextran sulfate sodium (DSS)-induced colitis, smooth muscle damage, and gut microbiota dysbiosis.

2. Line 106 and Line 109, "DSS-incduced" please spell correctly. 3. Line 268, "The fecal samples were collected between groups DSS+BBR and DSS", 16S rRNA gene sequencing study and Bioinformatics Analysis, how many groups of data? Two groups or three groups? 4. Please add the control group information in Fig 2 of figure legends 5. Please specify the linkages between reduced inflammation and smooth muscle in the colon.
Reviewer #2 (Comments for the Author): UC disease research is very common in human and animals, but there are few studies on cats. This study is of great significance to the development of cat research. As cats live together with humans, some diseases have good comparative medical significance. This is a new direction worth studying. In this paper, high-throughput sequencing was used to determine the bacterial changes in the intestinal tract of colitis. Berberine was found that could alleviate the intestinal flora disorder with dextran sodium sulfate-induced. The present study showed a new direction for the treatment of colitis. It was further verified that berberine could alleviate the inflammatory reaction and regulate the contraction of smooth muscle in intestinal. This paper was interesting. It has a certain guiding significance for the research of non-antibiotic therapy for the treatment of enteritis by relying on the regulation of microbiota. I recommend that this article be accepted for publication. 1.Please add the references in Line 62. 2.Line 67, "BBR had a significant effect on ulcerative colitis bioavailability", What is the bioavailability level? 3.Line 170, "signalling pathway", please spell correctly. 4.How about the successful rate of model construction? Is 18-day sufficient to construct the model? 5.There are still some shortcomings in English writing. It is suggested to modify and perfect it Staff Comments:

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